Lactobacillus crispatus capable of promoting female genital tract health and delaying genital tract immune aging

Abstract

The invention discloses lactobacillus crispatus for improving female genital tract microenvironment and immune aging and application of the lactobacillus crispatus, and belongs to the field of probiotics. The probiotic lactobacillus crispatus ProSci-38 disclosed by the invention can be used for effectively improving the symptoms such as pruritus vulvae, causalgia of vagina, abnormal vaginal secretion and the like of patients with vaginitis; mainly through the effect of intestinal tract-vaginal axis, the proportion of lactobacillus is increased, the proportion of harmful bacteria is reduced, the balance of vaginal flora is recovered, and the acidic environment of vagina is maintained; meanwhile, the inflammatory factor level can be reduced, the immune function of the vagina of a patient can be improved, so that the vagina environment is improved, the infection risk is reduced, and the preparation method has a great prospect in preparation of products for relieving and / or treating vaginitis.

Description

Technical Field

[0001] The invention relates to Lactobacillus crispatus for improving the microenvironment of female reproductive tract and delaying reproductive tract immune aging and applications thereof, and specifically relates to Lactobacillus crispatus ProSci-38 for improving the microenvironment of female reproductive tract and delaying reproductive tract immune aging and a preparation method and application thereof, belonging to the field of probiotics. Background Art

[0002] Vulvovaginitis is a common and frequently occurring gynecological disease with a high recurrence rate. It affects patients of all ages and affects their physical and mental health and quality of life. Normally, the microecology and immune function in the vagina are balanced. When the body is affected by its own or external factors, the natural defense function of the vagina is destroyed, the pH ratio is unbalanced, and the flora structure is unbalanced, increasing the risk of pathogen invasion and causing vaginal inflammation.

[0003] The vaginal microecological system is a unique system composed of the vagina's anatomical structure, microecological flora, local immune system and endocrine regulation function. The vaginal microbial flora is diverse and changes dynamically under the influence of various endogenous or exogenous factors. The dominant flora that resides the most in this balanced system is lactobacillus, accounting for about 50% to 80%. It can produce lactic acid to maintain a low pH value in the vagina, and stimulate the immune function of vaginal mucosal cells by secreting metabolites such as hydrogen peroxide, bacteriocins, and biosurfactants, inhibiting the survival and reproduction of some pathogenic bacteria and conditional pathogens, thereby maintaining the balance of the vaginal microecological environment. However, when the bacterial community structure is imbalanced, the pH ratio is abnormal, various pathogenic microorganisms invade, and the vaginal self-cleaning function is affected, various infectious diseases in the vagina will occur. The rational application rate of antibiotics is low and the recurrence rate is high. Its irrational use is an important factor in the reduction of lactobacillus, which may cause vaginal flora imbalance or secondary infection.

[0004] Therefore, the current treatment method that mainly aims to kill pathogenic microorganisms with antibiotics needs to be transformed into a treatment model that aims to increase probiotics, repair and restore the balance of the vaginal environment, rebuild the vaginal microecological system, and restore the defense function. It is urgent to develop a probiotic preparation that can improve the microenvironment of the female reproductive tract and delay reproductive tract immune aging, as a new treatment option for vaginitis. Summary of the invention

[0005] In view of the above-mentioned deficiencies in the prior art, the present invention provides Lactobacillus crispatus and its application for improving the microenvironment of the female reproductive tract and delaying reproductive tract immune aging, aiming to solve the current technical problem of lack of microbial agents for improving the microenvironment of the female reproductive tract and delaying reproductive tract immune aging.

[0006] The first technical solution provided by the present invention is a strain of Lactobacillus crispatus ProSci-38, which has been deposited in the General Microbiology Center of China National Microbiological Culture Collection Administration on June 20, 2024, and its deposit number is CGMCC No.31019.

[0007] Lactobacillus crispatus ProSci-38 was isolated from the female reproductive tract and was deposited at the General Microbiology Center of the China Microbiological Culture Collection Administration on June 20, 2024. The deposit address is No. 3, Yard No. 1, Beichen West Road, Chaoyang District, Beijing. Its microbial deposit number is CGMCC No.31019. The test result of the strain is survival. The strain has good tolerance to gastric acid and bile salts.

[0008] The second technical solution provided by the present invention is a microbial preparation containing the Lactobacillus crispatus ProSci-38 described in the first technical solution.

[0009] In some embodiments, the content of Lactobacillus crispatus ProSci-38 in the microbial preparation is not less than 1×10 6 Cells / mL or 1×10 6 Cells / g.

[0010] Furthermore, the content of the Lactobacillus crispatus ProSci-38 in the microbial preparation is not less than 1×10 6 CFU / mL or 1×10 6 CFU / g.

[0011] In certain embodiments, the microbial preparation comprises any one of the culture, fermentation product or lyophilized powder of the bacterial strain.

[0012] The third technical solution provided by the present invention is a postbiotic composition, which includes one or more of the fermentation supernatant, inactivated bacteria, bacterial lysate and fermentation broth of Lactobacillus crispatus ProSci-38 described in the first technical solution.

[0013] In certain embodiments, the postbiotic is a fermentation broth obtained by fermenting and culturing Lactobacillus crispatus ProSci-38.

[0014] In certain embodiments, the postbiotic is obtained by culturing Lactobacillus crispatus ProSci-38 to obtain a fermentation broth, and sterilizing and inactivating the fermentation broth at 85° C. for 15 minutes.

[0015] In certain embodiments, the postbiotic is obtained by inactivating the Lactobacillus crispatus ProSci-38 after fermentation and culture.

[0016] In certain embodiments, the postbiotic is obtained by high-pressure homogenization and membrane filtration of the fermentation broth of the Lactobacillus crispatus ProSci-38 fermentation culture to obtain a bacterial lysate.

[0017] In certain embodiments, the postbiotics can be dried into powder or used directly by various drying methods such as vacuum drying, spray drying, vacuum freeze drying, fluidized bed drying, etc.

[0018] The fourth technical solution provided by the present invention is a product containing the Lactobacillus crispatus ProSci-38 described in the first technical solution, the microbial preparation described in the second technical solution, or the postbiotic composition described in the third technical solution.

[0019] In certain embodiments, the product is a food, a medicine, or a hygiene product.

[0020] In certain embodiments, the food is a dairy product, a bean product, or a fruit and vegetable product produced by fermentation using the Lactobacillus crispatus ProSci-38 described in the first technical solution, or the microbial preparation described in the second technical solution.

[0021] In certain embodiments, the food is a beverage or snack containing the Lactobacillus crispatus ProSci-38 described in the first technical solution, or the microbial preparation of the second technical solution.

[0022] In certain embodiments, the medicine comprises the above-mentioned strain and / or preparation, and a pharmaceutically acceptable carrier.

[0023] Furthermore, the carrier includes one or more of fillers, binders, wetting agents, disintegrants, lubricants, and flavoring agents commonly used in medicine.

[0024] Furthermore, the dosage form of the drug includes granules, capsules, tablets, pills, suppositories or oral liquids.

[0025] Furthermore, the medicines include oral tablets and capsules with enteric coatings, oral liquids; vaginal suppositories, tablets, gelatin capsules, sprays, creams, and gels.

[0026] Furthermore, the sanitary products include sanitary wet wipes, sanitary napkins, sanitary pads, sanitary tampons, sanitary napkins, vaginal washes, and women's antibacterial / antibacterial washes.

[0027] In some embodiments, the content of Lactobacillus crispatus ProSci-38 in the product is not less than 1×106 Cells / mL or 1×10 6 Cells / g.

[0028] Furthermore, the content of the Lactobacillus crispatus ProSci-38 in the product is not less than 1×10 6 CFU / mL or 1×10 6 CFU / g.

[0029] The fifth technical solution provided by the present invention is the use of Lactobacillus crispatus ProSci-38 described in the first technical solution, the microbial preparation described in the second technical solution, or the postbiotic composition described in the third technical solution in the preparation of a product for preventing, alleviating and / or treating vaginitis.

[0030] The sixth technical solution provided by the present invention is the application of the Lactobacillus crispatus ProSci-38 described in the first technical solution, the microbial preparation described in the second technical solution, or the postbiotic composition described in the third technical solution, wherein the application includes any one of the following:

[0031] 1) Application in the preparation of products for preventing, alleviating and / or treating vulvar itching, vaginal burning pain and abnormal vaginal discharge caused by vaginitis;

[0032] 2) Application in the preparation of products for increasing beneficial bacteria in the vagina, reducing harmful bacteria, and regulating the balance of vaginal flora;

[0033] 3) Application in the preparation of products for maintaining the acidic environment of the vagina;

[0034] 4) Application in the preparation of products for reducing the level of related inflammatory factors and enhancing vaginal immunity.

[0035] In some embodiments, the content of Lactobacillus crispatus ProSci-38 in the product is not less than 1×10 6 Cells / mL or 1×10 6 Cells / g.

[0036] Furthermore, the content of the Lactobacillus crispatus ProSci-38 in the product is not less than 1×10 10 CFU / mL or 1×10 10 CFU / g.

[0037] In certain embodiments, the product is a food, a medicine, or a hygiene product.

[0038] Furthermore, the medicine comprises the above-mentioned strain and / or preparation, and a pharmaceutically acceptable carrier.

[0039] Furthermore, the carrier includes one or more of fillers, binders, wetting agents, disintegrants, lubricants, and flavoring agents commonly used in medicine.

[0040] Furthermore, the dosage form of the drug includes granules, capsules, tablets, pills, suppositories or oral liquids.

[0041] Furthermore, the medicines include oral enteric-coated tablets and capsules, oral liquids; vaginal suppositories, tablets, gelatin capsules, sprays, creams, and gels.

[0042] Furthermore, the sanitary products include sanitary wet wipes, sanitary napkins, sanitary pads, sanitary tampons, sanitary napkins, vaginal washes, and women's antibacterial / antibacterial washes.

[0043] The technical effects of the present invention are as follows:

[0044] The probiotic Lactobacillus crispatus ProSci-38 described in the present invention can effectively improve symptoms such as vulvar itching, vaginal burning pain, abnormal vaginal discharge, etc. in patients with vaginitis; it mainly increases the proportion of lactobacilli, reduces the proportion of harmful bacteria, restores the balance of vaginal flora, and maintains the acidic environment of the vagina through the action of the intestinal-vaginal axis; at the same time, it can reduce the level of inflammatory factors, improve the immune function of the patient's vagina, thereby improving the vaginal environment and reducing the risk of infection, and has great prospects in the preparation of products for relieving and / or treating vaginitis.

[0045] Biomaterial Deposit

[0046] The Lactobacillus crispatus ProSci-38 provided by the present invention is taxonomically named Lactobacillus crispatus, isolated from the female reproductive tract and deposited in the General Microbiology Center of the China Culture Collection Administration on June 20, 2024. The deposit address is No. 3, Yard No. 1, Beichen West Road, Chaoyang District, Beijing, and its microbial deposit number is CGMCC No. 31019. DETAILED DESCRIPTION

[0047] The preferred embodiments of the present invention are described below. It should be understood that the embodiments are for better explaining the present invention and are not used to limit the present invention.

[0048] If no specific experimental steps or conditions are specified in the examples, the conventional experimental steps or conditions described in the literature in the field can be used. If no manufacturer is specified for the reagents or instruments used, they are all conventional reagent products that can be purchased commercially. It should be noted that "Cells / mL or Cells / g" in the present invention is obtained by counting with a bacterial counting plate, and the results obtained are Total bacterial count including dead cells; "CFU / mL or CFU / g" refers to colony forming unit, that is, the total number of bacterial colonies per milliliter or per gram, which is obtained by the plate colony counting method.

[0049] Example 1 Isolation and Identification of Lactobacillus crispatus ProSci-38

[0050] Taking female reproductive tract secretions as samples, 0.5 mL of the sample was added to 5 mL of MRS liquid culture medium (commercially available), and anaerobically cultured at 37°C for 24 h for enrichment to obtain the enriched sample; 0.5 mL of the enriched sample was added to 4.5 mL of sterile saline to obtain a 10-1 dilution, and then 0.5 mL of the 10-1 dilution was added to 4.5 mL of saline to obtain a 10-2 dilution. In this way, 10-3, 10-4, 10-5, and 10-6 dilutions were obtained in sequence; 100 μL of the gradient dilutions were applied to MRS solid culture medium, 2 plates for each gradient of 10-4, 10-5, and 10-6, and anaerobically cultured at 37°C for 48 h to obtain colonies; colonies on MRS solid culture medium were selected according to the shape, size, edge, transparency, etc. of the colonies, and the colonies were picked with an inoculation loop and placed on the MRS solid culture medium Streak on the culture medium, and culture anaerobically at 37°C for 48 hours to obtain a purified single colony; pick the purified single colony and inoculate it into 5 mL of MRS liquid culture medium respectively, and culture it anaerobically at 37°C for 24 hours to obtain a bacterial solution; number each strain corresponding to each bacterial solution, and perform Gram staining, strain identification, physiological and biochemical experiments and genome identification analysis according to the steps recorded in the textbook "Microbiology" (edited by Shen Ping and Chen Xiangdong), and obtain a strain with typical characteristics of Lactobacillus crispatus, which is recorded as strain ProSci-38;

[0051] The process of bacterial species identification is as follows:

[0052] The strain ProSci-38 bacteria were taken, and the genome of the strain ProSci-38 was extracted using a bacterial genome extraction kit (commercially available). The 16S universal 27F / 1492R primer pair was used to amplify the extracted genome of the strain ProSci-38 as a template to obtain the 16S rRNA of the strain ProSci-38; the 16S rDNA of the strain ProSci-38 was compared with the nucleic acid sequence using the Blastn program of NCBI, and the results showed that this strain was Lactobacillus crispatus, and was named Lactobacillus crispatus ProSci-38. Lactobacillus crispatus ProSci-38 was deposited in the General Microbiology Center of China Microorganism Culture Collection Administration, with the deposit number CGMCC No.31019, the deposit address: No. 3, Yard 1, Beichen West Road, Chaoyang District, Beijing, Institute of Microbiology, Chinese Academy of Sciences, and the deposit date was June 20, 2024; the test result of the strain was survival.

[0053] Example 2

[0054] A method for preparing a microbial preparation (probiotic powder) comprises the following steps:

[0055] 1) Activation of bacterial strains: The strains stored at -40°C were inoculated into the corresponding liquid culture medium sterilized at 121°C for 15 minutes, and cultured anaerobically at 37°C for 18-24 hours. The activated bacterial strains were obtained by subculturing 1-2 times.

[0056] 2) Optimize the preparation of culture medium:

[0057] The components of the optimized culture medium were prepared in proportion, mixed evenly, adjusted to pH 6.5, and sterilized at 121°C for 15 min;

[0058] The composition of the modified MRS liquid culture medium is as follows: sucrose 23.7Kg, lactose 11.9Kg, soy peptone 15.0Kg, yeast powder 5.6Kg, yeast peptone 14.5Kg, Na2HPO3 23.1Kg, citric acid 1.4Kg, MgSO4·7H2O 0.8Kg, MnSO45H2O0.2Kg, Tween-80 1.4Kg, L-cysteine ​​hydrochloride 0.9Kg, and distilled water 1000L.

[0059] 3) Preparation of seed solution:

[0060] Each activated bacterial strain in step 1) is inoculated into the optimized culture medium prepared in step 2), and anaerobically cultured at 37° C. until the pH value reaches 4.5-4.8;

[0061] 4) Inoculation and fermentation:

[0062] The seed liquid of step 3) is inoculated into the optimized medium prepared in step 2) at 2‰, and fermented for 24 hours under controlled fermentation conditions;

[0063] The fermentation conditions were controlled as follows: constant temperature culture at 30°C in the early stage of fermentation, and natural fermentation until the pH reached 5.0; then the fermentation temperature was adjusted to constant temperature culture at 37°C, and the pH was controlled to remain at 6.0 to maintain anaerobic fermentation.

[0064] In the above process, pH is controlled by adding a neutralizing agent, wherein the neutralizing agent is NaOH.

[0065] 5) Stop fermentation

[0066] When the bacteria stopped producing acid, the fermentation was terminated and the high-density fermentation liquid of each strain was obtained. The number of live bacteria in the fermentation liquid reached 1×10 10 CFU / mL and above.

[0067] Whether the acid production has stopped can be determined by the fact that the pH no longer decreases and the addition of the neutralizer stops.

[0068] 6) Freeze drying

[0069] a. Bacteria concentration: The high-density fermentation broth is centrifuged at 12000g to concentrate the bacteria;

[0070] b. Add protective agent: Add 5 times the protective agent solution to the bacterial concentrate; the protective agent solution composition is as follows: 15.6Kg skim milk, 13.9Kg lactose, 1.2Kg vitamin C, 1.6Kg sodium glutamate, 1000L distilled water.

[0071] c. Drying: The above-mentioned bacterial suspension after adding the protective agent is freeze-dried to obtain freeze-dried bacterial powder, and the bacterial powders of each strain are mixed according to the ratio (ratio of the number of colony forming units) to control the total number of viable bacteria in the mixed bacterial powder to reach 1.0×10 11 CFU / g and above.

[0072] The total viable bacteria / cell count of the probiotic powder / probiotic postbiotic powder of Lactobacillus crispatus ProSci-38 prepared in this example is 1×10 6 CFU / Cells / g.

[0073] Example 3

[0074] This embodiment provides a postbiotic composition, wherein the components of the postbiotic composition include inactivated bacteria and metabolites of Lactobacillus crispatus ProSci-38; and the preparation method of the postbiotic composition is as follows:

[0075] Seed culture: 0.1 mL of Lactobacillus crispatus ProSci-38 (deposit number: CGMCC No. 31019) was inoculated into MRS liquid culture medium (purchased from Beijing Luqiao Technology Co., Ltd.), and cultured at 30° C. for 30 h to obtain ProSci-38 seed solution;

[0076] Fermentation: ProSci-38 seed liquid was inoculated into the fermentation medium, and the total viable count of Lactobacillus crispatus ProSci-38 was 2×10 6 CFU / mL, and then cultured at 30°C for 30 h to obtain the fermentation broth;

[0077] Post-treatment: sterilize the fermentation broth at 85°C for 15 min to obtain a sterilized and inactivated fermentation broth; spray dry the sterilized and inactivated fermentation broth to obtain a postbiotic composition (the total number of Lactobacillus crispatus ProSci-38 cells in the postbiotic composition is 1.0×10 11 Cells / g);

[0078] Wherein, the preparation method of fermentation medium is as follows:

[0079] The fermentation medium consists of 1% defatted soy flour, 8% defatted milk powder and the balance water in percentage by mass; the components of the fermentation medium are mixed in proportion, and the mixture is stirred at 58° C. for 15 minutes (i.e., stirred at 150 rpm) to obtain a liquid; the liquid is homogenized once at 58° C. at a primary pressure of 19 MPa and a secondary pressure of 5.0 MPa to obtain a homogenized liquid; the homogenized liquid is sterilized at 93° C. for 30 minutes to obtain a sterilized liquid; the sterilized liquid is cooled to 35° C. to obtain a fermentation medium.

[0080] Example 4

[0081] A probiotic / postbiotic milk powder comprises milk powder and the probiotic powder / postbiotic composition of Lactobacillus crispatus ProSci-38 prepared in Example 2 / Example 3. The specific preparation method is: firstly prepare milk powder by using a conventional milk powder preparation process, add the probiotic powder / postbiotic composition of Lactobacillus crispatus ProSci-38 to the prepared milk powder, and mix them evenly.

[0082] The number of viable bacteria / cells of a probiotic milk powder in this embodiment is not less than 1.0×10 6 CFU / Cells / g.

[0083] Example 5

[0084] A probiotic / postbiotic protein powder comprises protein powder and the probiotic powder / postbiotic composition of Lactobacillus crispatus ProSci-38 prepared in Example 2 / Example 3. The specific preparation method thereof is: firstly preparing protein powder by a conventional protein powder preparation process, adding the probiotic powder / postbiotic composition of Lactobacillus crispatus ProSci-38 to the prepared protein powder, and mixing them evenly.

[0085] The viable bacteria count / cell count of a probiotic protein powder in this embodiment is not less than 1.0×10 6 CFU / Cells / g.

[0086] Example 6

[0087] A probiotic / postbiotic crystal ball, comprising a crystal ball and a probiotic powder / postbiotic composition of Lactobacillus crispatus ProSci-38 prepared in Example 2 / Example 3. The specific preparation method is: a crystal ball of a specific composition is prepared according to the composition requirements of the crystal ball raw material, and the Lactobacillus crispatus ProSci-38 probiotic powder / postbiotic composition is added to the prepared crystal ball.

[0088] The number of live bacteria / cells of a probiotic / postbiotic crystal ball in this embodiment is not less than 1.0×10 6 CFU / Cells / g.

[0089] Example 7

[0090] A probiotic / postbiotic solid beverage comprises a solid beverage powder and a probiotic powder / postbiotic composition of Lactobacillus crispatus ProSci-38 prepared in Example 2 / Example 3. The specific preparation method is: a solid beverage powder of a specific composition is prepared according to the composition requirements of the solid beverage powder raw material, and the Lactobacillus crispatus ProSci-38 probiotic powder / postbiotic composition is added to the prepared solid beverage powder, and the mixture is mixed evenly.

[0091] The viable bacteria count / cell count of a probiotic / postbiotic solid beverage in this embodiment is not less than 1.0×10 6 CFU / Cells / g.

[0092] Example 8

[0093] A probiotic / postbiotic film candy comprises a film candy and a probiotic powder / postbiotic composition of Lactobacillus crispatus ProSci-38 prepared in Example 2 / Example 3. The specific preparation method is: mixing the raw materials of the film candy, adding the probiotic powder / postbiotic composition of Lactobacillus crispatus ProSci-38 during the raw material mixing process, mixing the raw materials and the probiotic powder / postbiotic composition of Lactobacillus crispatus ProSci-38 uniformly, and then using a conventional film candy process to prepare the probiotic / postbiotic composition film candy.

[0094] The number of viable bacteria / cells of a probiotic / postbiotic film candy in this embodiment is not less than 1.0×10 6 CFU / Cells / g.

[0095] Example 9

[0096] A probiotic / postbiotic tablet candy comprises a tablet candy and a probiotic powder / postbiotic composition of Lactobacillus crispatus ProSci-38 prepared in Example 2 / Example 3. The specific preparation method is: mixing raw materials of the tablet candy, adding the probiotic powder / postbiotic composition of Lactobacillus crispatus ProSci-38 during the raw material mixing process, uniformly mixing the raw materials and the probiotic powder / postbiotic composition of Lactobacillus crispatus ProSci-38, and then using a conventional tableting process to prepare the probiotic / postbiotic tablet candy.

[0097] The viable count of a probiotic / postbiotic tablet candy in this embodiment is not less than 1.0×10 6 CFU / Cells / g.

[0098] Example 10

[0099] A fermented milk is obtained by fermenting milk using milk as a raw material and Lactobacillus crispatus ProSci-38 as a fermentation bacterium. The preparation process of the fermented milk is a conventional process and will not be described in detail here.

[0100] The viable count of Lactobacillus crispatus ProSci-38 in the fermented milk obtained in this example should not be less than 1.0×10 6 CFU / g.

[0101] Embodiment 11

[0102] A sanitary napkin comprises a sanitary napkin and the probiotic postbiotic composition of Lactobacillus crispatus ProSci-38 prepared in Example 3.

[0103] The bacterial count of a sanitary napkin in this embodiment is not less than 1.0×10 6 Cells / g.

[0104] Example 12

[0105] A sanitary wet wipe comprises the sanitary wet wipe and the probiotic postbiotic composition of Lactobacillus crispatus ProSci-38 prepared in Example 3.

[0106] The bacterial count of a sanitary wet wipe in this embodiment is not less than 1.0×10 6 Cells / g.

[0107] Example 13

[0108] A private parts care liquid comprises the private parts care liquid and the probiotic postbiotic composition of Lactobacillus crispatus ProSci-38 prepared in Example 3.

[0109] The bacterial count of a private parts care solution in this embodiment is not less than 1.0×10 6 Cells / g.

[0110] Embodiment 14

[0111] An underwear washing liquid comprises the underwear washing liquid and the probiotic postbiotic composition of Lactobacillus crispatus ProSci-38 prepared in Example 3.

[0112] The bacterial count of an underwear washing liquid in this embodiment is not less than 1.0×10 6 Cells / g.

[0113] Embodiment 15

[0114] A private area care gel comprises the private area care gel and the probiotic postbiotic composition of Lactobacillus crispatus ProSci-38 prepared in Example 3.

[0115] The bacterial count of a private care gel in this embodiment is not less than 1.0×10 6 Cells / g.

[0116] Example 16

[0117] A private parts care suppository comprises a private parts care suppository and a probiotic postbiotic composition of Lactobacillus crispatus ProSci-38 prepared in Example 3.

[0118] The bacterial count of a private part care suppository in this embodiment is not less than 1.0×10 6 Cells / g.

[0119] Embodiment 17

[0120] A private parts care spray comprises a private parts care spray and a probiotic postbiotic composition of Lactobacillus crispatus ProSci-38 prepared in Example 3.

[0121] The bacterial count of a private care spray in this embodiment is not less than 1.0×10 6 Cells / g.

[0122] Embodiment 18

[0123] The strain Lactobacillus crispatus ProSci-38 contained in the probiotic powder prepared in Example 2 was provided by Beijing Ketuo Hengtong Biotechnology Co., Ltd.

[0124] A total of 168 patients with bacterial vaginitis who visited the Inner Mongolia People's Hospital from January 2024 to June 2024 were selected and randomly divided into three groups. Experimental group: 56 cases, average age (36.43±5.08) years old, 45 pregnant cases, 11 non-pregnant cases; observation group: 56 cases, average age (35.98±4.55) years old, 42 pregnant cases, 14 non-pregnant cases; and control group: 56 cases, average age (36.15±5.23) years old, 44 pregnant cases, 12 non-pregnant cases. There was no significant difference in the general data of the three groups (P>0.05), which was comparable. Informed consent was obtained from the patients in this study. During the treatment period, the three groups of patients were required to pay attention to genital hygiene, maintain healthy living habits, eat a reasonable diet, and avoid sexual life.

[0125] Inclusion criteria: Meet the diagnostic criteria for bacterial vaginosis; have no allergic reaction to the study drugs; and have high patient compliance.

[0126] Exclusion criteria: patients with a history of antibiotics and vaginal topical medications within the past month; patients who experienced adverse events during the study; patients with incomplete follow-up data or who dropped out midway; patients with AIDS, other underlying diseases, pregnancy, and unmarried.

[0127] Treatment method: The control group was given 1 tablet of metronidazole suppository vaginally, once a day, 7 days as a course of treatment, and the drug was discontinued after one course of treatment; the observation group was given 1 tablet of Luna Probiotics (1.2×10 10 CFU, Lactobacillus rhamnosus CA15, Lactobacillus rhamnosus PLV68, Lactobacillus acidophilus PLV107 and Lactobacillus crispatus PLV103 for 28 days; the experimental group took Lactobacillus crispatus ProSci-38 probiotic powder (1.2×10 10 CFU / day) for a total of 28 days.

[0128] Clinical efficacy: The clinical efficacy was evaluated with reference to the "Clinical Disease Diagnosis Based on Treatment Cure and Improvement Standards". Cured: The patient's vulvar itching, vaginal burning and other symptoms disappeared, the vaginal discharge returned to normal, and the sialidase was negative; markedly effective: The patient's above clinical symptoms improved by more than 60%, the vaginal discharge improved significantly, and the sialidase was weakly positive; effective: The patient's above clinical symptoms improved by 20-60%, the vaginal discharge improved, and the sialidase was positive; ineffective: The patient's symptoms and signs did not improve, and the sialidase was positive. Total effective rate = (cured + markedly effective + effective) ÷ total number of cases × 100%.

[0129] Observation indicators: ① Distribution of vaginal flora: beneficial bacteria are mainly lactobacilli, and harmful bacteria include enterobacteria, bacteroides, mycoplasma, and Candida albicans. Vaginal secretion samples were collected from patients before and after treatment for bacterial culture, and the bacteria on the culture medium were identified. ② Vaginal microecology: The proportion of patients with vaginal pH ≤ 4.5 and the proportion of patients with lactobacilli dominance before and after treatment in the three groups were compared. ③ Vaginal immune function indicators: including interleukin-2 (IL-2), interleukin-10 (IL-10), interleukin-12 (IL-12), tumor necrosis factor-α (TNF-α) and gamma-interferon (IFN-γ). Enzyme-linked immunosorbent assay was used to measure vaginal secretion samples before and after treatment.

[0130] Table 1

[0131]

[0132] The efficacy of the three groups of patients at the end of the trial was significantly different. The total effective rate of the control group was 87.5%, and the total effective rates of the experimental group and the observation group were 98.2% and 92.7%, respectively. The proportion of cured patients was significantly higher than that of the control group, as shown in Table 1. This shows that oral probiotics can effectively improve symptoms such as vulvar itching, vaginal burning pain, and abnormal vaginal discharge in patients, and is an effective auxiliary treatment for vaginitis.

[0133] Table 2

[0134]

[0135] Before treatment, there was no significant difference in the distribution of vaginal flora types among the three groups of patients. After 7 days of treatment, the distribution of Lactobacillus increased in the three groups, while the distribution of Bacteroides, Enterobacter, Candida albicans, and Mycoplasma decreased, and the improvement effects of the experimental group and the observation group were better than those of the control group. After 28 days of treatment, the distribution of Lactobacillus continued to increase in the experimental group and the observation group, and the distribution of Bacteroides, Enterobacter, Candida albicans, and Mycoplasma decreased again, while all indicators of the control group rebounded. Antibiotics inhibit bacterial growth and prevent the growth of pathogenic microorganisms by hindering the synthesis of bacterial deoxyribonucleotides during the treatment of vaginitis, thereby improving the vaginal environment, but antibiotic treatment alone cannot make up for the reduction of Lactobacillus. Oral probiotics mainly increase the proportion of Lactobacillus, reduce the proportion of harmful bacteria, and restore the balance of vaginal flora through the action of the intestinal-vaginal axis, thereby improving the vaginal environment and reducing the risk of infection.

[0136] Table 3

[0137]

[0138]

[0139] The higher the proportion of patients with vaginal pH ≤ 4.5 and the proportion of Lactobacillus dominant, the better the vaginal microecology of the patients. After 7 days of treatment, the proportion of patients with vaginal pH ≤ 4.5 and the proportion of Lactobacillus dominant in the three groups increased significantly, indicating that the vaginal environment of the patients improved. After 28 days of treatment, these two indicators in the experimental group and the observation group continued to improve, especially in the experimental group, which was as high as 100% and 98.4%, while the control group had no such effect. This shows that the intake of probiotic Lactobacillus crispatus ProSci-38 has a positive effect on the flora of patients with vaginitis, reduces harmful bacteria in the vagina, and promotes the growth of beneficial bacteria. It can produce lactic acid, thereby maintaining the acidic environment of the vagina to prevent the excessive growth of harmful microorganisms.

[0140] Table 4

[0141]

[0142] Vaginitis may be related to abnormal local immune function, and vaginal aging will also accelerate the aging of the reproductive system. Before treatment, the levels of IL-2, IL-10, IL-12, TNF-α and IFN-γ in patients were all at a high level, indicating that the body had vaginitis. After 28 days of treatment, the levels of IL-2, IL-10, IL-12, TNF-α and IFN-γ in patients in the experimental group and the observation group were significantly reduced, indicating that cytokines play a certain role in vaginal immunity in patients with bacterial vaginitis. It shows that the intake of probiotics can improve the immune function of patients with vaginitis, reduce vaginal inflammatory reactions, and thus promote the recovery of patients.

[0143] Embodiment 19

[0144] A total of 68 patients with bacterial vaginitis who visited the Inner Mongolia People's Hospital from May 2024 to June 2024 were selected and randomly divided into three groups, namely, postbiotic group A (23 cases, average age (34.90±1.13) years old), postbiotic group B (20 cases, average age (35.70±1.77)) and control group (25 cases, average age (35.05±2.43)). The subjects cleaned their hands and vulva with warm water every night before use during the menstrual period, took a gynecological antibacterial gel product, lay on their backs with their hips slightly raised, slowly inserted the gel catheter containing the gel liquid into the deep vagina, and pushed it into the vulva with a push rod, and maintained the posture for about 15 minutes. It can be used continuously for 14 days, one per night. During this period, family members were advised not to have sexual intercourse or other vaginal invasive operations, and other antibiotics, probiotics and other drugs should not be used during treatment. The difference is that the postbiotic group A used gynecological antibacterial gel with probiotic postbiotic powder containing Lactobacillus crispatus ProSci-38, the postbiotic group B used gynecological antibacterial gel with probiotic postbiotic powder containing Lactobacillus crispatus NM194-4, and the control group used Renhe gynecological gel.

[0145] Observation index: ① Clinical efficacy: refer to the method in Example 1. ② Duration of symptoms and signs: Symptoms and signs observation indicators include vaginal itching, vaginal pain, and abnormal secretions. The changes in disease symptoms of the three groups of patients before and after treatment were observed, and the score was set to 0-10 points, where 0 was asymptomatic and 10 was severe symptoms. The score was positively correlated with the symptom manifestation. ③ Vaginal microecological index: Vaginal specimens of patients were collected before and after treatment, and Gram staining microscopy was performed to examine the bacterial density of level II to III (level II bacterial density: 10 to 99 bacteria per oil immersion field; level III bacterial density: ≥100 bacteria per oil immersion field), the dominant bacterial flora was Lactobacillus (normal standard is +++~++++, where 6 to 30 per field of view is +++, and >30 per field of view is ++++), and the incidence of vaginal secretion pH value ≤4.5.

[0146] Table 5

[0147]

[0148] The clinical efficacy of postbiotic group A was the highest at 95.7%, and the proportion of cured patients was significantly higher than that of the other two groups, as shown in Table 5. This indicates that the gynecological gel with the addition of Lactobacillus crispatus ProSci-38 probiotic postbiotic powder can effectively improve the symptoms of vulvar itching, vaginal burning pain, abnormal vaginal discharge, etc.

[0149] Table 6

[0150]

[0151] After treatment, the symptom scores of the three groups of patients were lower than before treatment, and the duration of symptoms was reduced, and the effect of postbiotic group A was the best, as shown in Table 6. It further shows that the use of gynecological gel with added Lactobacillus crispatus ProSci-38 probiotic postbiotic powder can treat or relieve symptoms such as vaginal itching, vaginal pain, abnormal vaginal discharge, etc., and shorten the duration of symptoms.

[0152] Table 7

[0153]

[0154]

[0155] After treatment, the vaginal microecological indicators of the three groups of patients improved. The incidence of Lactobacillus as the dominant flora, flora density II-III, and vaginal secretion pH ≤ 4.5 in the postbiotic A group was higher than that in the postbiotic B group and the control group. This indicates that the use of gynecological gel with added Lactobacillus crispatus ProSci-38 probiotic postbiotic powder can effectively regulate vaginal pH and maintain vaginal microecological balance. Lactic acid bacteria play a role in the vagina by competitive adhesion, gaining a dominant position, and then inhibiting the attachment of pathogens, thereby achieving the antibacterial purpose.

[0156] In summary, the intake of probiotic Lactobacillus crispatus ProSci-38 can effectively improve the patient's clinical symptoms such as vulvar itching, vaginal burning pain, abnormal vaginal discharge, etc.; and regulate the balance of vaginal flora by lowering the vaginal pH value and increasing the proportion of lactobacillus; at the same time, it can reduce the level of inflammatory factors, improve the patient's vaginal immune function, and reduce vaginal inflammatory response, thereby promoting the patient's recovery.

[0157] Although the present invention has been disclosed as above in the form of a preferred embodiment, it is not intended to limit the present invention. Anyone familiar with this technology can make various changes and modifications without departing from the spirit and scope of the present invention. Therefore, the scope of protection of the present invention should be based on the definition of the claims.

Claims

1. A strain of Lactobacillus crispatus ProSci-38, characterized in that: The Lactobacillus crispatus ProSci-38 was deposited in the General Microbiology Center of China Microorganism Culture Collection Administration on June 20, 2024, and its deposit number is CGMCC No.31019.

2. A microbial preparation containing the Lactobacillus crispatus ProSci-38 according to claim 1.

3. The microbial preparation according to claim 2, characterized in that The content of Lactobacillus crispatus ProSci-38 in the microbial preparation is not less than 1×10 6 Cells / mL or 1×10 6 Cells / g.

4. A postbiotic composition, characterized in that The postbiotic composition comprises one or more of the fermentation supernatant, inactivated bacteria, bacterial lysate and fermentation broth of the Lactobacillus crispatus ProSci-38 according to claim 1.

5. A product containing the Lactobacillus crispatus ProSci-38 according to claim 1, the microbial preparation according to claim 2 or 3, or the postbiotic composition according to claim 4.

6. The product according to claim 5, characterized in that The product is food, medicine or sanitary product.

7. The product according to claim 5, characterized in that The content of Lactobacillus crispatus ProSci-38 in the product is not less than 1×10 6 Cells / mL or 1×10 6 Cells / g.

8. Use of the Lactobacillus crispatus ProSci-38 according to claim 1, the microbial preparation according to claim 2 or 3, or the postbiotic composition according to claim 4 in the preparation of a medicine for preventing, alleviating and / or treating vaginitis.

9. The use according to claim 8, characterized in that: The application also includes: application in the preparation of medicines for preventing, alleviating and / or treating vulvar itching, vaginal burning pain, and abnormal vaginal discharge caused by vaginitis.

10. Use of the Lactobacillus crispatus ProSci-38 according to claim 1, the microbial preparation according to claim 2 or 3, or the postbiotic composition according to claim 4 in the preparation of medicines, characterized in that: The medicines include any of the following: 1) Drugs that increase beneficial bacteria in the vagina, reduce harmful bacteria, and regulate the balance of vaginal flora; 2) Drugs that maintain the acidic environment of the vagina; 3) Drugs that reduce the levels of related inflammatory factors IL-2, IL-10, IL-12, TNF-α and / or IFN-γ and enhance vaginal immunity.

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